IFNγ ELISpot assay on splenocytes of mice vaccinated two times with TMV-DOST vaccine.

Abstract

<p>Splenocytes were isolated from 2 mice per group on D65 after intranasal vaccination and incubated on IFNγ antibody coated plates at a final concentration of 10⁵ cells/well. Whole protein antigen stimulants were added at 20 μg/mL, and plates incubated at 37°C in a 5% CO₂, 99% humidity incubator for ~36h. IFNγ secreting cells were detected and developed using Mouse IFNγELISpot Ready-SET-Go! Kit, developed with AEC, and read on an AID ELISpot plate reader. IFNγ secretion from spleen cells stimulated with DnaK (<b>A</b>.) OmpA (<b>B.</b>) SucB (<b>C.</b>) and Tul4 (<b>D</b>.) are compared to no stimulation (no stim), or PBS vaccinated mice (Norm). Values were adjusted positive IFNγ spots per 10⁶ cells, and groups that show significant augmentation with adjuvants compared to no adjuvant are shown (*p<0.05, **p<0.001). Means are presented ± standard deviation, and p values were calculated by non-parametric t-test. Results are representative of two experiments.</p